Fig. 2

HIF-1 alpha overexpression does not modify the ability of MSCs to inhibit T-cell activation. T cell-enriched peripheral blood cells were stained with carboxyfluoroscein succinimidyl ester (CFSE) and activated with anti-CD3/anti-CD28 monoclonal antibodies in the presence or absence of mesenchymal stem cells (MSCs; light grey) and hypoxia-inducible factor (HIF)-MSCs (dark grey). T cells (TC) were cultured in media alone as a negative control. After 4 days, expression of CD25 and proliferation of T-cell subsets was determined by flow cytometry. A representative experiment (a) and the mean ± SD of three independent experiments (b) are shown. T cells were gated according to forward/side scatter characteristics and expression of CD3, CD4, and CD8. c Supernatants of cultures were collected after 4 days and assayed for interferon (IFN)-gamma production. Data are presented as mean ± SD of three independent experiments. One-way ANOVA, *p ≤ 0.05, GFP-MSCs and HIF-MSCs versus no MSCs. GFP green fluorescent protein